A chemiluminescence immunoassay for rapid detection of classical swine fever virus E2 antibodies in pig serum samples.

Abstract

The high performance of chemiluminescence immunoassays (CLIAs) in diagnosis has been gradually recognized in recent years, but their application in the diagnosis of classical swine fever (CSF) has not been reported. Here, a recombinant E2 (rE2) protein and a peroxidase-conjugated monoclonal antibody (MAb G5) were used to develop a competition-based chemiluminescence immunoassay (cCLIA) for rapid and accurate detection of E2-specific antibodies in pig serum. To evaluate the feasibility of cCLIA in the diagnosis of CSF, we developed a competition-based enzyme-linked immunosorbent assay (cELISA) as a control. Under the optimum test conditions, cCLIA showed a higher signal-to-noise ratio than that of the control cELISA. The best signal-to-noise ratios of cCLIA and cELISA were 70 and 17, respectively. Then, the diagnostic performance of the two assays was compared by examining a panel of pig serum samples (n=285) with a confirmed status, and cCLIA showed higher diagnostic sensitivity (Dn) and diagnostic specificity (Dp) values than those of cELISA. The Dn and Dp of cCLIA were 97.49% and 96.08%, respectively, and those of cELISA were 93.97% and 94.12%, respectively. Furthermore, cCLIA can provide results within 20min, whereas the control cELISA requires at least 1hr. According to these findings, the newly developed cCLIA has potential application in the diagnosis of CSF and offers an alternative approach for efficient and rapid detection of E2-specific antibodies.